May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
Suppression of ICAM-1 in Murine Retina by Plasmid Small Interfering RNAs
Author Affiliations & Notes
  • Y. Hirano
    Dept of Ophthalmology and Vis Sci, Nagoya City Univ Med Sch, Nagoya, Japan
  • E. Sakurai
    Dept of Ophthalmology and Vis Sci, Nagoya City Univ Med Sch, Nagoya, Japan
  • A. Matsubara
    Dept of Ophthalmology and Vis Sci, Nagoya City Univ Med Sch, Nagoya, Japan
  • Y. Ogura
    Dept of Ophthalmology and Vis Sci, Nagoya City Univ Med Sch, Nagoya, Japan
  • Footnotes
    Commercial Relationships  Y. Hirano, None; E. Sakurai, None; A. Matsubara, None; Y. Ogura, None.
  • Footnotes
    Support  None.
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 5343. doi:https://doi.org/
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      Y. Hirano, E. Sakurai, A. Matsubara, Y. Ogura; Suppression of ICAM-1 in Murine Retina by Plasmid Small Interfering RNAs. Invest. Ophthalmol. Vis. Sci. 2008;49(13):5343. doi: https://doi.org/.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : Leukocyte plays critical role for many kinds of ocular disease such as uveitis, diabetic retinopathy, choroidal neovascularization and so on. Intercellular adhesion molecule -1(ICAM-1) is essential for this migratory process in retinal vessels or leukocyte. Suggesting, control of ICAM-1 expression lead to strategy of therapy for such disease. Small interfering ribonucleic acids (siRNAs) are efficient and specific modulators of endogenous gene expression. This study describes the application of siRNAs to suppress ICAM-1 expression on the surface of mice retinal pigmented epithelial (RPE) cells using a hydrodynamics-based transfection technique.

Methods: : ICAM-1 specific plasmid siRNAs designed from the mice gene sequence were transfected into the retina using a hydrodynamics-based transfection technique in vivo. We evaluated the presence of plasmid siRNA in the mice retina by green fluorescent protein (GFP) expression plasmid. Upregulation of ICAM-1 was induced by streptozotocin (STZ)-induced diabetes mellitus mouse model. ICAM-1 expression was analyzed by immunohistochemistry, ELISA and flow cytometry.

Results: : After the GFP expression plasmid administration, the fluorescence of GFP was found in the each layer of the mice retina histologically. ICAM-1 expression in control mice was 0.5±0.1ng/ml in the retina. Seven days after the STZ administration, ICAM-1 expression was 2.1±0.3ng/ml, and 1.2±0.1ng/ml in the plasmid ICAM-1 siRNA transfected group (n=12, P<0.05 vs STZ retina).

Conclusions: : These findings suggest that ICAM-1 expression can be effectively silenced by plasmid siRNAs on mice retina in vivo. This new technology may render novel therapeutic concepts to reduce retinal neovascular disease by inhibition of leukocyte infiltration.

Keywords: gene transfer/gene therapy • microscopy: light/fluorescence/immunohistochemistry • retina 
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